Genetic Modification of Adenovirus Vectors for Bone Cancer Treatment
Session Number
BIO 14
Advisor(s)
Tong-Chuan He, University of Chicago
Discipline
Biology
Start Date
17-4-2025 10:45 AM
End Date
17-4-2025 11:00 AM
Abstract
This paper aims to examine the complex interplay of bone morphogenetic protein 9 (BMP-9) with osteogenesis, osteosarcoma, and its many other functions in the arena of research and translation into clinical therapeutics. Six recombinant adenovirus lines containing different heparin-binding (HB) domains that can increase the osteogenic activity of BMP-9 were constructed and then separated into two groups. One group of the six lines had a red fluorescent protein (RFP) cassette inserted and the other group had a green fluorescent protein (GFP) cassette inserted. The fluorescence helped with precise titer determination. Multiple rounds of cell infection were conducted by adding the recombinant adenovirus lines to human embryonic kidney (HEK) 293 cells to create a high-titer stock and establish a virus bank that can be used for virus production. It was discovered that the titer with higher cell density and GFP showed more consistent infection of the cells within seven days than that with lower cell density and RFP. The titer with higher cell density consistently infected about 20% of cells in 24 hours and 30% after 36 hours with replating of the cells. This work demonstrated that the parameters needed, such as cell density, can be optimized for virus preservation and titer stability. In the future, stable genes of interest, such as HB domain and BMP-9, can be produced using the recombinant adenoviruses studied above, and then be effectively and efficiently delivered into mammalian cells. HB domain has shown to be able to increase the osteogenic activity of BMP-9, which can slow down the growth of osteosarcoma. Therefore, recombinant adenoviruses with HB domain and BMP-9 are very promising for future osteosarcoma treatments.
Genetic Modification of Adenovirus Vectors for Bone Cancer Treatment
This paper aims to examine the complex interplay of bone morphogenetic protein 9 (BMP-9) with osteogenesis, osteosarcoma, and its many other functions in the arena of research and translation into clinical therapeutics. Six recombinant adenovirus lines containing different heparin-binding (HB) domains that can increase the osteogenic activity of BMP-9 were constructed and then separated into two groups. One group of the six lines had a red fluorescent protein (RFP) cassette inserted and the other group had a green fluorescent protein (GFP) cassette inserted. The fluorescence helped with precise titer determination. Multiple rounds of cell infection were conducted by adding the recombinant adenovirus lines to human embryonic kidney (HEK) 293 cells to create a high-titer stock and establish a virus bank that can be used for virus production. It was discovered that the titer with higher cell density and GFP showed more consistent infection of the cells within seven days than that with lower cell density and RFP. The titer with higher cell density consistently infected about 20% of cells in 24 hours and 30% after 36 hours with replating of the cells. This work demonstrated that the parameters needed, such as cell density, can be optimized for virus preservation and titer stability. In the future, stable genes of interest, such as HB domain and BMP-9, can be produced using the recombinant adenoviruses studied above, and then be effectively and efficiently delivered into mammalian cells. HB domain has shown to be able to increase the osteogenic activity of BMP-9, which can slow down the growth of osteosarcoma. Therefore, recombinant adenoviruses with HB domain and BMP-9 are very promising for future osteosarcoma treatments.